scholarly journals The hedgehog-patched signaling pathway and function in the mammalian ovary: a novel role for hedgehog proteins in stimulating proliferation and steroidogenesis of theca cells

Reproduction ◽  
2009 ◽  
Vol 138 (2) ◽  
pp. 329-339 ◽  
Author(s):  
Leon J Spicer ◽  
Satoko Sudo ◽  
Pauline Y Aad ◽  
Lora Shuo Wang ◽  
Sang-Young Chun ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Mrna Abundance ◽  
Rt Pcr ◽  
Theca Cells ◽  
Qrt Pcr ◽  
Hedgehog Proteins ◽  
Patched 1 ◽  
And Function ◽  
First Time

The expression of hedgehog (Hh) genes, their receptor, and the co-receptor in mice, rat, and bovine ovaries were investigated. RT-PCR of ovarian transcripts in mice showed amplification of transcripts for Indian (Ihh) and desert (Dhh) Hh, patched 1 (Ptch1), and smoothened (Smo) genes. Semi-quantitative RT-PCR and northern blot analyses showed that whole ovarianIhhandDhhtranscripts decreased 4–24 h after hCG versus 0–48 h after pregnant mares serum gonadotrophin treatment in mice, whereas mousePtch1andSmotranscripts were expressed throughout the gonadotropin treatments. Quantitative real-time RT-PCR (qRT-PCR) revealed that the expression of the Hh-patched signaling system withIhhmRNA abundance in granulosa cells was greater, whereasSmoandPtch1mRNA abundance was less in theca cells of small versus large follicles of cattle. In cultured rat and bovine theca-interstitial cells, qRT-PCR analyses revealed that the abundance ofGli1andPtch1mRNAs were increased (P<0.05) with sonic hedgehog (SHH) treatment. Additional studies using cultured bovine theca cells indicated that SHH induces proliferation and androstenedione production. IGF1 decreasedIhhmRNA abundance in bovine granulosa cells. The expression and regulation ofIhhtranscripts in granulosa cells andPtch1mRNA in theca cells suggest a potential paracrine role of this system in bovine follicular development. This study illustrates for the first time Hh activation of Gli1 transcriptional factor in theca cells and its stimulation of theca cell proliferation and androgen biosynthesis.

scholarly journals Localization of gonadotrophin-releasing hormone I, bradykinin and their receptors in the ovaries of non-mammalian vertebrates

Reproduction ◽  
2007 ◽  
Vol 133 (5) ◽  
pp. 969-981 ◽  
Author(s):  
Padmasana Singh ◽  
Amitabh Krishna ◽  
Rajagopala Sridaran
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Cell Types ◽  
Physiological Significance ◽  
Rt Pcr ◽  
Releasing Hormone ◽  
Theca Cells ◽  
Gonadotrophin Releasing Hormone

GnRH I and its receptors have been demonstrated in the ovaries of various vertebrates, but their physiological significance in reproductive cascade is fragmentary. Bradykinin is a potent GnRH stimulator in the hypothalamus. In the present study, the presence of GnRH I and its receptor, and bradykinin and its receptor in the ovaries of non-mammalian vertebrates were investigated to understand their physiological significance. GnRH I immunoreactivity in the ovaries of fish, frog, reptile and bird were mainly found in the oocyte of early growing follicles and granulosa cells and theca cells of previtellogenic follicles. Vitellogenic follicles showed mild GnRH immunoreactivity. GnRH I-receptor and bradykinin were localized in the same cell types of the ovaries of these vertebrates. The presence of GnRH I, GnRH I-receptor and bradykinin in the ovaries of these vertebrates was confirmed by immunoblotting. The presence of GnRH I mRNA was demonstrated in the ovary of vertebrates using RT-PCR. The ovaries of reptiles and birds showed significantly higher intensity of immunoreactivity for GnRH I-receptor as compared with the fish and amphibian. This may have a correlation with the higher yolk content in the ovary of reptile and bird. These results suggest the possibility of GnRH I and bradykinin as important regulators of follicular development and vitellogenesis in the vertebrate ovary.

scholarly journals Expression and effect of NAMPT (visfatin) on progesterone secretion in hen granulosa cells

Reproduction ◽  
2015 ◽  
Vol 150 (1) ◽  
pp. 53-63 ◽  
Author(s):  
Mélodie Diot ◽  
Maxime Reverchon ◽  
Christelle Ramé ◽  
Yannick Baumard ◽  
Joëlle Dupont
Keyword(s):  
Granulosa Cells ◽  
Specific Inhibitor ◽  
Follicular Development ◽  
Rt Pcr ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Theca Cells ◽  
Protein Levels ◽  
Progesterone Secretion ◽  
Ovulatory Follicles

In mammals, nicotinamide phosphoribosyltransferase (NAMPT) is an adipokine produced by adipose tissue that is found in intracellular and extracellular compartments. The intracellular form of NAMPT is a nicotinamide phosphoribosyltransferase, whereas the extracellular form is considered an adipokine. In humans, NAMPT regulates energy metabolism and reproductive functions, such as ovarian steroidogenesis. To date, no study has investigated the role of NAMPT in hen ovaries. We investigated whether NAMPT is present in hen ovarian follicles and its role in granulosa cells. Using RT-PCR, western blotting and immunocytochemistry, we detected mRNA transcripts and proteins related to NAMPT in theca and granulosa cells from pre-ovulatory follicles. Using RT-PCR, we demonstrated that mRNA NAMPT levels were higher in granulosa cells than they were in theca cells and that during follicle development, theca cell levels decreased, whereas levels remained unchanged in granulosa cells. NAMPT protein quantities were significantly higher in theca cells than they were in granulosa cells, but they were unchanged during follicular development. Plasma NAMPT levels, as determined by ELISA and immunoblotting, were significantly lower in adult hens than they were in juveniles. In vitro, treatment with human recombinant NAMPT (100 ng/ml, 48 h) halved basal and IGF1-induced progesterone secretion, and this was associated with a reduction in STAR and HSD3B protein levels and MAPK3/1 phosphorylation levels in granulosa cells. These effects were abolished by the addition of FK866, a specific inhibitor of NAMPT enzymatic activity. Moreover, NAMPT had no effect on granulosa cell proliferation. In conclusion, NAMPT is present in hen ovarian cells and inhibits progesterone production in granulosa cells.

scholarly journals Regulation and action of fibroblast growth factor 17 in bovine follicles

2009 ◽  
Vol 202 (3) ◽  
pp. 347-353 ◽  
Author(s):  
M F Machado ◽  
V M Portela ◽  
C A Price ◽  
I B Costa ◽  
P Ripamonte ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Granulosa Cells ◽  
Cumulus Cells ◽  
Mrna Abundance ◽  
Fibroblast Growth ◽  
Theca Cells ◽  
Progesterone Secretion ◽  
Igf I

Fibroblast growth factor 17 (FGF17) is a member of the FGF8 subfamily that appears to be relevant to folliculogenesis and oogenesis, as the prototype member FGF8 is an oocyte-derived protein that signals to cumulus cells. FGF8 has structural and receptor-binding similarities to FGF17, whose expression in the ovary has not been reported. In this study, we demonstrate localization of FGF17 protein to the oocyte of preantral follicles, and to the oocyte and granulosa cells of antral follicles. Real-time PCR demonstrated the presence of mRNA in oocytes and, to a lesser extent, in granulosa and theca cells. FGF17 mRNA abundance was low in granulosa and theca cells from healthy follicles and increased significantly in atretic follicles. Addition of FSH or IGF-I to granulosa cells in vitro decreased FGF17 mRNA abundance, and treatment with FGF17 inhibited estradiol and progesterone secretion from granulosa cells in relation to control cultures without these additives. We conclude that FGF17 is a potential mediator of granulosa cell differentiation.

scholarly journals Expression and Localization of Secreted Frizzled-Related Protein-4 in the Rodent Ovary: Evidence for Selective Up-Regulation in Luteinized Granulosa Cells

Endocrinology ◽  
2003 ◽  
Vol 144 (10) ◽  
pp. 4597-4606 ◽  
Author(s):  
Minnie Hsieh ◽  
Sabine M. Mulders ◽  
Robert R. Friis ◽  
Arun Dharmarajan ◽  
JoAnne S. Richards
Keyword(s):  
Chorionic Gonadotropin ◽  
Granulosa Cells ◽  
Human Chorionic Gonadotropin ◽  
Follicular Development ◽  
Corpora Lutea ◽  
Related Protein ◽  
Rich Domain ◽  
And Function ◽  
Periovulatory Follicles

Secreted frizzled-related protein-4 (sFRP-4) belongs to a family of soluble proteins that have a Frizzled-like cysteine-rich domain and function as modulators of Wnt-Frizzled (Fz) signals. As several Wnts and Fz are expressed at defined stages of follicular development in rodent ovaries, these studies were undertaken to evaluate the hormone-regulated expression and localization of sFRP-4. In the mouse ovary, the expression of sFRP-4 mRNA was up-regulated in granulosa cells of large antral follicles after human chorionic gonadotropin administration and was also elevated in corpora lutea, as determined by RT-PCR and in situ hybridization analyses. In hypophysectomized rat ovaries, sFRP-4 expression was similarly induced by human chorionic gonadotropin and further up-regulated by PRL. PRL also stimulated the secretion of sFRP-4 protein from luteinized rat granulosa cells in culture. Therefore, regulation of sFRP-4 by LH and PRL may be important for modulating Fz-1, which is known to be expressed in periovulatory follicles, and Wnt-4/Fz-4, which are expressed in corpora lutea.

010. Oocyte control of granulosa cell development and function

2005 ◽  
Vol 17 (9) ◽  
pp. 66
Author(s):  
J. J. Eppig ◽  
K. Sugiura
Keyword(s):  
Energy Source ◽  
Amino Acid ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Paracrine Factors ◽  
Subtraction Hybridization ◽  
Junctional Communication ◽  
And Function

Oocytes orchestrate the rate of follicular development and the patterns of gene expression by granulosa cells (GCs). There are two populations of GCs in large antral follicles: mural granulosa cells (MGCs) that line the ovarian follicle wall, and cumulus cells (CCs) closely associated with the oocyte. Subtraction hybridization was used to find transcripts more highly expressed in CCs than MGCs. Among the genes expressed more highly in CCs was one encoding an amino acid transporter (Slc38a3). Slc38a3 mRNA was not detected in oocytes. Expression of Slc38a3 mRNA was reduced in the CCs after removal of the oocyte and restored by co-culturing CCs with fully grown oocytes (FGOs). Alanine is one of the amino acids transported by SLC38A3. This amino acid is poorly transported across the oocyte plasma membrane, but gains access to the oocyte from the cumulus cells via gap junctional communication. Alanine transport into cumulus cells was promoted by paracrine factors secreted by FGOs, but not by growing oocytes (GOs) from preantral follicles. Thus FGOs promote the transport of alanine into CCs, and this amino acid is then passed on to the oocyte via gap junctions. Transcripts encoding enzymes in the glycolytic pathway were also more highly expressed in CCs than MGCs. FGOs, but not GOs, promote elevated expression of some of these transcripts. Likewise, FGOs promote both glycolysis and oxidative phosphorylation by isolated CCs and MGCs. Oocytes do not effectively utilize glucose as an energy source, and oocytes require the presence of CCs to resume meiosis when glucose is the only energy source present. In contrast, oocytes can resume meiosis in the absence of CCs when pyruvate is the sole energy source. Thus oocytes apparently promote glycolysis by their companion granulosa cells to provide energy for their own development. In addition, this may be one way that oocytes coordinate their development with that of follicular somatic components. Supported by Grants HD23839 and HD44416 from the NICHD.

Real-time RT-PCR quantification of pregnancy-associated plasma protein-A mRNA abundance in bovine granulosa and theca cells: Effects of hormones in vitro

2006 ◽  
Vol 31 (4) ◽  
pp. 357-372 ◽  
Author(s):  
Pauline Y. Aad ◽  
Justin L. Voge ◽  
Consuelo A. Santiago ◽  
Jerry R. Malayer ◽  
Leon J. Spicer
Keyword(s):  
Real Time ◽  
Plasma Protein ◽  
Protein A ◽  
Mrna Abundance ◽  
Rt Pcr ◽  
Theca Cells

Immunohistochemical localization of steroidogenic enzymes and comparison with hormone production during follicle development in the pig

1999 ◽  
Vol 11 (6) ◽  
pp. 337 ◽  
Author(s):  
Ellen M. Shores ◽  
Morag G. Hunter
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Staining Intensity ◽  
Immunohistochemical Localization ◽  
Vital Role ◽  
Steroidogenic Enzymes ◽  
Hormone Production ◽  
Theca Cells ◽  
Enzyme Distribution ◽  
Follicle Size

The steroidogenic enzymes, P450 aromatase (P450 arom ) and P450 17a-hydroxylase (P450 17a ), were precisely located within the healthy porcine follicle by immunohistochemistry. Enzyme distribution was examined throughout follicular development during natural oestrous cycles (n = 14 gilts) and was compared with steroid production by healthy whole and theca-only follicles. All follicles 2 mm or more in diameter were either fixed for immunohistochemistry (n = 380 of which 197 were assessed as healthy) or incubated as whole (n = 110) or theca-only (n = 110) follicles to measure steroidogenesis. P450 17a was confined to the theca layer. The number of positive cells and staining intensity increased with follicle size. P450 arom was consistently detected in the granulosa layer of follicles measuring 6 mm or more in diameter and those cells furthest from the antrum were most strongly stained. P450arom was also detected in the theca layer of these large follicles. Whole and theca-only follicles produced oestradiol and androstenedione, and the levels of both hormones increased with follicle size (P<0.001). Whole follicles produced more oestradiol (P<0.001), but less androstenedione (P = 0.01) than theca-only follicles of the same size. Although granulosa cells contained P450 arom and synthesized oestradiol, only theca cells contained P450 17a. Theca cells therefore provided granulosa cells with androgen substrate. In addition, theca cells possessed P450 arom , making them capable of independent oestradiol production, which may be required to trigger the LH surge. This study confirms the vital role of theca cells in follicular steroidogenesis in the pig.

scholarly journals The Water Permeability Channels Aquaporins 1–4 Are Differentially Expressed in Granulosa and Theca Cells of the Preovulatory Follicle during Precise Stages of Human Ovulation

2011 ◽  
Vol 96 (4) ◽  
pp. 1021-1028 ◽  
Author(s):  
Asgeir Thoroddsen ◽  
Pernilla Dahm-Kähler ◽  
Anna Karin Lind ◽  
Birgitta Weijdegård ◽  
Bernhard Lindenthal ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
University Hospital ◽  
Mrna Levels ◽  
Theca Cells ◽  
Cell Compartments ◽  
Obstetrics And Gynaecology ◽  
Early Rise ◽  
Laparoscopic Sterilization ◽  
First Time ◽  
Ovulatory Process

Abstract Context: Changes in vascular permeability and expansion of the fluid-filled antrum are major events in the LH-induced ovulatory process. Objectives: Our objective was to investigate the presence and expression levels of aquaporins (AQPs) in the granulosa and theca cell compartments of the follicle during defined phases of human ovulation. Design and Setting: We conducted a prospective experimental study at the Department of Obstetrics and Gynaecology at a university hospital. Participants: Twenty-eight women underwent laparoscopic sterilization and at the same time follicle retrieval at four periovulatory phases. Main Outcome Measures: mRNA levels of AQP1-4 were measured in separated granulosa and theca cells from preovulatory phase, early ovulatory (EO) phase, late ovulatory phase, and postovulatory phase. Immunohistochemistry was done for AQP1-4 in intact human follicles. Results: All four AQPs were expressed in both the theca and granulosa cells during ovulation. In granulosa cells, AQP1 levels increased in the late ovulatory and postovulatory phases. Expression of AQP2-3 followed a similar pattern with a marked increase in the EO phase, whereas AQP4 levels decreased from preovulatory to the EO phase. The presence of AQP1-4 in the human follicle was verified by immunohistochemistry. Conclusions: The results show for the first time the presence of AQP1-4 in human follicles during ovulation. The marked early rise in expression of AQP2 and AQP3 suggests a role during the process leading to follicular rupture, and the late rise of AQP1 suggests a role in corpus luteum formation.

scholarly journals Oocyte control of ovarian follicular development and function in mammals

Reproduction ◽  
2001 ◽  
pp. 829-838 ◽  
Author(s):  
JJ Eppig
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Primordial Follicle ◽  
Dominant Factor ◽  
Ovarian Follicular Development ◽  
New Perspective ◽  
Regulatory Loop ◽  
Sphere Of Influence ◽  
And Function

A new perspective on ovarian follicular development has emerged over the last decade. Whereas the oocyte was previously considered only a passive recipient of developmental signals from oocyte-associated granulosa cells, it is now clear that communication between oocytes and granulosa cells is bidirectional. A complex interplay of regulatory factors governs the development of both types of cell. This interplay is essential not only for oocyte development but also for follicular development, beginning with the initial assembly of the primordial follicle and continuing throughout ovulation. The existence of an oocyte-granulosa cell regulatory loop, essential for normal follicular differentiation as well as for the production of an oocyte competent to undergo fertilization and embryogenesis, is proposed. Although gonadotrophins are essential for driving the differentiation of granulosa cell phenotypes, within its sphere of influence, the oocyte is probably the dominant factor determining the direction of differentiation and the function of the granulosa cells associated with it.

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